Enabling Efficient, Fast, Biocompatible Exosome Separation via Acoustofluidics

通过声流控技术实现高效、快速、生物相容性的外泌体分离

基本信息

批准号：
10171868
负责人：
Tony Jun Huang
金额：
$ 42.96万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10171868
关键词：
Acoustics Address Albumins Alzheimer&apos s Disease Antigen Presentation Benchmarking Biological Biological Markers Biological Process Biomedical Research Blood Blood Circulation Cell Culture Techniques Cell physiology Cells Centrifugation Cerebrospinal Fluid Chylomicrons Clinical Communities Containment of Biohazards Contracts DNA Devices Diagnosis Diagnostic Dimensions Disease Drug Delivery Systems Freezing Glass Health High Density Lipoproteins Human Resources Hybrids IDL lipoproteins Immunoglobulins In Vitro Industry Standard Kidney Diseases Lipoproteins Liquid substance Liver diseases Low-Density Lipoproteins Malignant Neoplasms Messenger RNA Methods MicroRNAs Microfluidics Molecular Molecular Profiling Monitor Morphology Neurodegenerative Disorders Parkinson Disease Patients Performance Phase Placenta Diseases Procedures Prognosis Property Proteins Recovery Research Research Personnel Saliva Sampling Series Serum Proteins Side Sorting - Cell Movement Speed Techniques Technology Therapeutic Time Training Transducers Urine Very low density lipoprotein Vesicle base biological research biomarker discovery biomaterial compatibility cell type circulating biomarkers clinical application cost density design exosome extracellular vesicles improved innovation interactive feedback intercellular communication microfluidic technology nanoscale nanosized non-Native operation particle physical property point of care preservation prognostic tool

项目摘要

Abstract Exosomes are nanosized extracellular vesicles that contain biomolecules (DNA, mRNA, miRNA, and other functional proteins) from their cell of origin. Exosomes are secreted from nearly all cell types, and as a result, they are found in most biological fluids, including blood, saliva, urine, and cerebrospinal fluid. Over the past decade, the transfer of exosomal biomolecules to recipient cells has been implicated in a variety of biological processes. Consequently, exosomes have increasingly been the focus of many studies in biomedical research. Due to their distinct molecular signatures, exosomes have been identified as a potentially transformative circulating biomarker for the diagnosis and prognosis of multiple diseases, including cancer, neurodegenerative diseases (i.e., Parkinson’s and Alzheimer’s), as well as diseases of the kidney, liver, and placenta. In addition to diagnostic applications, exosomes are an ideal drug delivery system in many therapeutic applications. While the versatility of exosomes renders them an excellent candidate for a variety of biomedical applications, difficulties in the consistent, effective isolation of exosomes have greatly limited their utility. Current approaches for exosome isolation involve lengthy procedures, require highly trained personnel, suffer from low repeatability, low yield, low purity, and/or low post-sorting exosome integrity. As a result, there exists a critical need in the research communities for a simple, rapid, efficient, and biocompatible approach for isolating exosomes form biological fluids or in vitro cell culture. In this R01 project, we will address this unmet need by developing an acoustofluidic (i.e., the fusion of acoustics and microfluidics) platform for high-purity, high-yield, high-biocompatibility, automated exosome isolation. The proposed acoustofluidic technology will have the following features: 1) Automated exosome processing which reduces operator-to-operator variability and enables simple, consistent isolation results with improved biohazard containment; 2) Reduces the amount of time necessary to go from biofluid (e.g., 1 mL undiluted blood) to isolated exosomes (<5 min processing time vs ~8 hrs processing time with alternative technologies); 3) Higher exosome recovery rate (>90%) in comparison to benchmark technologies (5‒25%); 4) Greater exosome purity (>80%) in comparison to benchmark technologies (~33%); 5) Less contamination from other circulating factors, including non-native serum proteins (e.g., albumin and immunoglobulin) and particles with similar sizes, including various types of lipoproteins; 6) Low-cost and point- of-care design; and 7) ability to handle both large and small sample volumes (maximum sample volume: ~30 mL; minimum sample volume: ~10 µL), which is extremely challenging with existing approaches. With these unique features, the proposed acoustofluidic technology has the potential to greatly simplify and expedite workflows in exosome-related biomedical research and aid in the discovery of new exosomal biomarkers.

抽象的外泌体是纳米大小的细胞外囊泡，含有生物分子（DNA、mRNA、miRNA 和其他几乎所有细胞类型都会分泌外泌体，因此，它们存在于大多数生物体液中，包括血液、唾液、尿液和脑脊液。十年来，外泌体生物分子向受体细胞的转移涉及多种生物学经过检查，外泌体越来越成为生物医学研究中许多研究的焦点。由于其独特的分子特征，外泌体已被认为是一种潜在的变革性药物用于多种疾病诊断和预后的循环生物标志物，包括癌症、神经退行性疾病疾病（即帕金森病和阿尔茨海默病），以及肾脏、肝脏和胎盘疾病。在诊断应用中，外泌体是许多治疗应用中理想的药物递送系统。外泌体的多功能性使其成为各种生物医学应用、困难的优秀候选者外泌体的一致、有效的分离极大地限制了它们目前的方法。外泌体分离涉及冗长的程序，需要训练有素的人员，重复性低，低产量、低纯度和/或低分选后外泌体完整性因此，研究存在迫切需求。社区寻求一种简单、快速、有效且生物相容的方法来从生物中分离外泌体在这个 R01 项目中，我们将通过开发声流体来解决这一未满足的需求。（即声学和微流控的融合）高纯度、高产量、高生物相容性的平台，所提出的声流体技术将具有以下特点：1) 自动化外泌体处理减少了操作员之间的差异，并实现简单、一致隔离结果改善了生物危害遏制；2) 减少了隔离所需的时间；生物流体（例如，1 mL 未稀释的血液）到分离的外泌体（<5 分钟处理时间与 ~8 小时处理时间 3) 与基准相比，外泌体回收率更高（>90%）技术 (5-25%)；4) 与基准技术 (~33%) 相比，外泌体纯度更高 (>80%)；其他循环因子的污染较少，包括非天然血清蛋白（例如白蛋白和免疫球蛋白）和类似尺寸的颗粒，包括各种类型的脂蛋白； 6）低成本和点- 护理设计；7) 能够处理大样本量和小样本量（最大样本量：~30 mL；最小样品体积：~10 µL），这对现有方法来说极具挑战性。独特的功能，所提出的声流技术有可能大大简化和加速外泌体相关生物医学研究的工作流程并有助于发现新的外泌体生物标志物。